CN110845634B - Polysaccharide extract in oil-tea camellia seed cake and extraction method thereof - Google Patents
Polysaccharide extract in oil-tea camellia seed cake and extraction method thereof Download PDFInfo
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Abstract
The invention relates to a polysaccharide extract in oil-tea camellia seed cake and an extraction method thereof, wherein the extraction method comprises the following steps: 1) adding a eutectic solvent into the crushed oil-tea camellia cakes, and performing ultrasonic centrifugation to obtain a polysaccharide crude extract; 2) adding a copolymer into the crude extract to construct an aqueous two-phase system, extracting polysaccharide to an upper phase rich in the copolymer, wherein the lower eutectic solvent can be recycled; 3) separating the copolymer phase, heating the copolymer containing polysaccharide to construct a thermal separation double-aqueous-phase system again to obtain a polysaccharide-containing aqueous phase and a copolymer phase, and recovering the copolymer; 4) drying the aqueous phase to obtain a primarily purified polysaccharide extract. The invention adopts a novel green solvent eutectic solvent as an extracting agent; polysaccharide is purified by adopting a copolymer/eutectic solvent aqueous two-phase system, polysaccharide separation and copolymer recycling are realized by heating, and further the extraction cost is reduced.
Description
Technical Field
The invention relates to the technical field of medicine extraction, and particularly relates to a polysaccharide extract in a camellia oleifera abel cake and an extraction method thereof.
Background
At present, the traditional methods for extracting polysaccharide from oil-tea camellia cakes comprise a water extraction method, an enzyme extraction method and an alkali extraction method. Wherein, the water extraction method is simple and has lower cost but lower efficiency. The enzyme extraction requires complicated extraction conditions, increasing the cost. The alkali extraction method has high efficiency, but the strong alkalinity also causes certain pollution.
Disclosure of Invention
Based on the above, the method for extracting the polysaccharide from the camellia oleifera abel cake is provided aiming at the technical defects of the method for extracting the polysaccharide from the camellia oleifera abel cake in the prior art.
A method for extracting polysaccharide from oil tea cakes comprises the following steps:
1) adding a eutectic solvent and water into the crushed oil tea cakes, and performing ultrasonic centrifugation to obtain a polysaccharide crude extract;
2) adding a copolymer into the obtained crude extract to form a double aqueous phase system, standing for phase separation to obtain a copolymer phase containing polysaccharide and a eutectic solvent phase, and recovering the eutectic solvent;
3) heating to enable the copolymer phase containing the polysaccharide to form a double-water-phase system again, performing phase separation to obtain a water phase containing the polysaccharide and the copolymer phase, and recovering the copolymer;
4) drying the obtained water phase to obtain polysaccharide extract.
In some embodiments, the copolymer comprises ethylene oxide-propylene oxide; the molecular weight of the ethylene oxide-propylene oxide is 2000-3000; the concentration of the ethylene oxide-propylene oxide is 30-70 wt%.
In some embodiments, the eutectic solvent comprises a hydrogen bond acceptor and a hydrogen bond donor, and the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is (1-5): (1-3), wherein the hydrogen bond acceptor comprises at least one of glycine, L (-) -proline, choline chloride and betaine, and the hydrogen bond donor comprises at least one of L (+) -lactic acid, DL-malic acid, glycerol, D-sorbitol, urea, ethylene glycol and citric acid.
In some embodiments, the amount of water added in the eutectic solvent is 10 to 90 wt%.
In some embodiments, the solid-to-liquid ratio of the camellia oleifera cake to the eutectic solvent is 1: (10-50).
In some embodiments, in step 1), the temperature of the ultrasound is 40 to 80 ℃; the ultrasonic time is 30-70 min.
In some embodiments, in the step 1), after the eutectic solvent is added to the crushed oil tea cake, the pH of the mixed solution is adjusted to 1-13.
In some embodiments, in step 2), after the copolymer is added to the obtained crude extract, the mixture is heated to 20 to 60 ℃.
In some embodiments, in step 3), the temperature of heating is 70 to 80 ℃.
The invention also provides a polysaccharide extract in the camellia oleifera abel cake, which is prepared by the extraction method in any one of the embodiments.
The method adopts a green eutectic solvent as an extracting agent to extract polysaccharide from the oil-tea camellia dead cake to obtain a polysaccharide crude extract, then adds a copolymer to construct a copolymer/eutectic solvent aqueous two-phase system, and the polysaccharide is extracted to a copolymer phase. Separating the two phases, heating the copolymer phase to form a thermally separated aqueous two-phase system again, extracting the polysaccharide into an aqueous phase, and drying the water after the two phases are separated to obtain a purified polysaccharide extract. Meanwhile, the copolymer can be recycled. Compared with the prior art, the invention has the following beneficial effects; a novel green solvent eutectic solvent is adopted as an extractant; polysaccharide is purified by adopting a copolymer/eutectic solvent aqueous two-phase system, polysaccharide separation and copolymer recycling are realized by heating, and further the extraction cost is reduced.
Drawings
Fig. 1 is a schematic flow chart of the method for extracting polysaccharide extract from oil-tea camellia seed cake disclosed by the invention.
Detailed Description
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, but rather should be construed as broadly as the present invention is capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
Example 1
As shown in fig. 1, a method for extracting polysaccharides from oil tea cakes comprises the following steps:
1) adding 0.1g of crushed and sieved oil tea cake powder into a 10mL centrifuge tube, and then adding 3.0mL of DES solution (containing 30 wt% of water) with the solid-liquid ratio of 1: 30; placing the centrifugal tube in an ultrasonic cleaning instrument for ultrasonic assisted extraction, wherein the ultrasonic temperature is 55 ℃, and the ultrasonic extraction time is 35 min; then centrifugally separating out supernatant to obtain polysaccharide crude extract;
2) adding 1.0mL of polysaccharide crude extract into a 10mL centrifuge tube, then adding EOPO copolymer, stirring the mixture uniformly, and standing to separate phases; wherein, the upper phase is an EOPO phase rich in polysaccharide, the lower phase is a DES phase, and the eutectic solvent is recovered;
3) heating to 70 deg.C in water bath to phase-separate the polysaccharide-containing copolymer phase to obtain polysaccharide-containing water phase and copolymer phase, and recovering the copolymer;
4) drying the obtained water phase to obtain polysaccharide extract.
5) And (3) analyzing and determining the content of the polysaccharide by adopting an anthrone-sulfuric acid method. The polysaccharide was measured on an ELISA plate using glucose as a standard. The detection wavelength is 625nm, the temperature is room temperature, the standard curve of polysaccharide determination is that y is 5.5893x-0.0051, and the correlation coefficient is R2Linear range 0.01-0.06mg/mL 0.9991.
Example 2
The reaction was carried out in the same manner as in example 1 except that different eutectic solvents shown in Table 1 below by the numbers 1 to 17 were used. In addition, this example also used water as the extractant to form the control group. The reaction results are shown in Table 1.
TABLE 1
| DES numbering | Hydrogen bond acceptors | Hydrogen bond donors | Molar ratio of | Extraction yield (mg/g) |
| DES-1 | Glycine | L (+) -lactic acid | 1:3 | 68.27 |
| DES-2 | L (-) -proline | DL-malic acid | 1:1 | 93.32 |
| DES-3 | L (-) -proline | Glycerol | 5:2 | 63.35 |
| DES-4 | L (-) -proline | L (+) -lactic acid | 1:1 | 59.33 |
| DES-5 | L (-) -proline | D-sorbitol | 1:1 | 56.20 |
| DES-6 | L (-) -proline | Glycerol | 1:2 | 57.54 |
| DES-7 | L (-) -proline | Urea | 1:2 | 106.29 |
| DES-8 | Choline chloride | L (+) -lactic acid | 1:1 | 97.79 |
| DES-9 | Choline chloride | Glycerol | 1:2 | 108.98 |
| DES-10 | Choline chloride | Ethylene glycol | 1:2 | 121.95 |
| DES-11 | Choline chloride | Urea | 1:2 | 102.27 |
| DES-12 | Choline chloride | Citric acid | 2:1 | 87.06 |
| DES-13 | Choline chloride | DL-malic acid | 1:1 | 73.19 |
| DES-14 | Betaine | L (+) -lactic acid | 1:1 | 85.72 |
| DES-15 | Betaine | Glycerol | 1:2 | 60.67 |
| DES-16 | Betaine | Urea | 1:2 | 88.85 |
| DES-17 | Betaine | Ethylene glycol | 1:2 | 102.71 |
| Water (W) | 80.35 |
As can be seen from table 1, when the hydrogen bond acceptor is choline chloride and the hydrogen bond donor is ethylene glycol in the eutectic solvent, and the molar ratio of the two is 1:2, the extraction yield of the polysaccharide is the highest.
Example 3
The same process as that of example 1 is carried out by adopting different reaction conditions, wherein the temperature of ultrasonic extraction is 50 ℃, the extraction time is 30min, and the addition amount of water is 10-90 wt%. The reaction results are shown in Table 2.
TABLE 2
As can be seen from Table 2, the extraction yield of the polysaccharide was the highest when the amount of water added was 30 wt%.
Example 4
The same procedure as in example 1 was followed, but under different reaction conditions, with 30 wt.% of water being added to the DES, the extraction temperature being 50 ℃ and the extraction time being 30 min. The reaction results are shown in Table 3.
TABLE 3
| Solid-to-liquid ratio | Extraction yield (mg/g) |
| 1：10 | 107.41 |
| 1：20 | 121.50 |
| 1：30 | 130.89 |
| 1：40 | 129.55 |
| 1：50 | 128.88 |
As can be seen from table 3, the extraction yield of polysaccharide was the highest when the solid-to-liquid ratio of camellia oleifera cake powder to DES was 1: 30.
Example 5
The procedure was as in example 1, but with different reaction conditions. Wherein, 30 wt% of water is added into DES, the solid-to-liquid ratio is 1:30, and the extraction time is 30 min. The reaction results are shown in Table 4.
TABLE 4
| Ultrasonic temperature (. degree. C.) | Extraction yield (mg/g) |
| 40 | 114.79 |
| 50 | 130.22 |
| 60 | 133.58 |
| 70 | 122.84 |
| 80 | 111.44 |
As can be seen from Table 4, the extraction yield of polysaccharide was the highest when the sonication temperature was 60 ℃.
Example 6
The procedure was as in example 1, but with different reaction conditions. Wherein, 30 wt% of water is added into DES, the solid-liquid ratio is 1:30, and the extraction temperature is 60 ℃. The reaction results are shown in Table 5.
TABLE 5
| Ultrasonic time (min) | Extraction yield (mg/g) |
| 30 | 126.20 |
| 40 | 132.23 |
| 50 | 139.61 |
| 60 | 135.59 |
| 70 | 128.21 |
As can be seen from Table 5, the extraction yield of polysaccharide was the highest when the sonication time was 50 min.
Example 7
The same procedure as in example 1 was followed, but with different reaction conditions, i.e., the pH of the mixture of the camellia oleifera cake and the DES solution was adjusted. Wherein 30 wt% of water is added into DES, the solid-liquid ratio is 1:30, the extraction temperature is 60 ℃, and the extraction time is 50 min. The reaction results are shown in Table 6.
TABLE 6
| pH of the system | Extraction yield (mg/g) |
| 1 | 123.51 |
| 3 | 138.27 |
| 5 | 145.65 |
| 7 | 134.25 |
| 9 | 132.23 |
| 11 | 124.18 |
| 13 | 65.14 |
As can be seen from Table 6, the polysaccharide extraction yield was the highest when the pH in the system was 5.
Example 8
The procedure was as in example 1, but with different reaction conditions. Wherein the concentration of the ethylene oxide-propylene oxide (EOPO) is 60 wt%, the temperature of the aqueous two-phase extraction is 30 ℃, and the pH value is 5. The reaction results are shown in Table 7.
TABLE 7
| EOPO molecular weight | EOPO phase extraction efficiency (%) |
| 2000 | 83.23 |
| 2500 | 84.92 |
| 3000 | 82.17 |
As can be seen from Table 7, the extraction efficiency was the highest when the molecular weight of EOPO was 2500.
Example 9
The procedure was as in example 1, but with different reaction conditions. Wherein, the molecular weight of EOPO is 2500, the aqueous two-phase extraction temperature is 30 ℃, and the pH value is 5. The reaction results are shown in Table 8.
TABLE 8
| EOPO concentration | EOPO phase extraction efficiency (%) |
| 30 | 80.66 |
| 40 | 81.09 |
| 50 | 82.32 |
| 60 | 85.4 |
| 70 | 80.45 |
As can be seen from Table 8, the extraction efficiency reached the highest when the concentration of EOPO was 60 wt%.
Example 10
The procedure was as in example 1, but with different reaction conditions. Wherein the molecular weight of EOPO is 2500, the concentration is 60 wt%, and the pH is 5. The reaction results are shown in Table 9.
TABLE 9
| Aqueous two-phase extraction temperature (. degree. C.) | EOPO phase extraction efficiency (%) |
| 20 | 80.18 |
| 30 | 85.16 |
| 40 | 86.92 |
| 50 | 81.55 |
| 60 | 82.98 |
As can be seen from Table 9, the extraction efficiency was the highest when the aqueous two-phase extraction temperature was 30 ℃.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (8)
1. The method for extracting polysaccharide from the oil tea dried cake is characterized by comprising the following steps:
1) adding a eutectic solvent solution into the crushed oil-tea camellia seed cake, performing ultrasonic assisted extraction, and then performing centrifugal separation to obtain a supernatant to obtain a polysaccharide crude extract; the addition amount of water in the eutectic solvent solution is 10-90 wt%;
2) adding a copolymer into the obtained crude extract to form a double aqueous phase system, standing for phase separation to obtain a copolymer phase containing polysaccharide and a eutectic solvent phase, and recovering the eutectic solvent; the copolymer is an ethylene oxide-propylene oxide copolymer; the molecular weight of the ethylene oxide-propylene oxide copolymer is 2000-3000; the concentration of the ethylene oxide-propylene oxide copolymer is 30-70 wt%;
3) heating to enable the copolymer phase containing the polysaccharide to form a double-water-phase system again, performing phase separation to obtain a water phase containing the polysaccharide and the copolymer phase, and recovering the copolymer;
4) drying the obtained water phase to obtain polysaccharide extract.
2. The extraction method according to claim 1, wherein the eutectic solvent comprises a hydrogen bond acceptor and a hydrogen bond donor, and the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is (1-5): (1-3), wherein the hydrogen bond acceptor comprises at least one of glycine, L (-) -proline, choline chloride and betaine, and the hydrogen bond donor comprises at least one of L (+) -lactic acid, DL-malic acid, glycerol, D-sorbitol, urea, ethylene glycol and citric acid.
3. The extraction method according to claim 1, wherein the solid-to-liquid ratio of the oil tea cake to the eutectic solvent solution is 1: (10-50).
4. The extraction method according to claim 1, wherein in the step 1), the temperature of the ultrasonic wave is 40-80 ℃; the ultrasonic time is 30-70 min.
5. The extraction method according to claim 1, wherein in the step 1), after the eutectic solvent solution is added to the crushed oil tea cake, the pH of the mixed solution is adjusted to 1-13.
6. The extraction method according to claim 1, wherein in the step 2), the copolymer is added to the obtained crude extract, and the mixture is heated to 20 to 60 ℃ after separation and purification.
7. The extraction method according to claim 1, wherein the heating temperature in step 3) is 70 to 80 ℃.
8. Polysaccharide extract in oil tea cake, characterized in that it is prepared by the extraction method of any one of claims 1 to 7.
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Application publication date: 20200228 Assignee: Shenzhen Xihan health Co.,Ltd. Assignor: INSTITUTE OF BAST FIBER CROPS, CHINESE ACADEMY OF AGRICULTURAL SCIENCES Contract record no.: X2023980032543 Denomination of invention: Polysaccharide extract from camellia oleifera dry cake and its extraction method Granted publication date: 20210903 License type: Common License Record date: 20230227 |